The study was conducted to assess the influence of different agro-ecological zones and cropping seasons on the incidence and severity of maize streak disease (MSD) in Ghana. Field surveys were conducted in two districts each of the coastal savannah, forest, and transitional agro-ecological zones of the Volta region to assess the incidence and severity of MSD in farmers' fields, during the 2014 minor and 2015 major cropping seasons. The disease assessment was carried out in 12 fields from each district, and for each field on both maize plants growing under tree shade and in open parts of the field. The plants were scored for disease severity based on a 1-5 visual scale (1=no infection and 5=very severe infection). Percentage total N, available P, exchangeable K, organic matter levels and pH were determined from soil samples collected from each of the fields surveyed. Incidence and severity of MSD on maize plants growing under tree shade and in open parts of the fields were significantly higher (P<0.05) in the transition zone than in the forest and coastal savannah eco-zones during the 2014 and 2015 cropping seasons. Mean incidence and severity of MSD on maize plants growing under tree shade were significantly higher than those in the open part of the fields in both cropping seasons. The levels of MSD incidence and severity recorded in 2014 were significantly higher (P<0.05) than 2015. Low levels of soil total N, available P, exchangeable K and organic carbon were detected across the three agro-ecological zones. Incidence and severity scores of MSD were significantly and negatively correlated with soil total N, available P, exchangeable K and organic matter (P<0.01). In conclusion, MSD incidence and severity vary with the cropping seasons and agro-ecological zones and are partly affected by low soil fertility levels and tree shades.